Preparation and purification of DNA from insects for AFLP analysis.
Analysis of amplified fragment length polymorphism (AFLP) has the potential to become a powerful new DNA fingerprinting technique for studying genetic relationships and genetic diversity in arthropods. Since DNA of high quality is a crucial prerequisite for AFLP analysis we evaluated the applicability of six protocols (one fast and four complex methods with phenol-chloroform treatments as well as one CTAB-based method) for extracting DNA from insect material and three additional DNA purification steps. The most rapid DNA isolation method did not produce DNA suitable for AFLP analysis. Among four complex methods tested, two protocols resulted in comparatively low yields of DNA that was therefore not used as template for AFLP analysis. The other two complex methods with phenol treatments and a CTAB-based DNA extraction protocol provided DNA suitable for AFLP assay. An additional purification of the DNA using spermine precipitation revealed a few extra bands in an AFLP gel that were masked in unpurified DNA. Therefore spermine precipitation is recommended for AFLP templates.
Reineke
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A
Institute of Phytomedicine, University of Hohenheim, Stuttgart, Germany. areineke@uni-hohenheim.de
Karlovsky
P
P
Zebitz
C P
CP
D000818:Animals; D004247:DNA; D016172:DNA Fingerprinting / Q000379:methods*; D007313:Insecta / Q000235:genetics*; D013698:Templates, Genetic
D004247:DNA